Naegleria fowleri from Pakistan Has Type-2 Genotype

Muhammad Aurongzeb; Yasmeen Rashid; Syed Habib  Ahmed Naqvi; Ambrina  Khatoon; Sadia  Abdul Haq; Muhammad Kamran Azim; Imdad  Kaleem; Shahid  Bashir

doi:10.18502/ijpa.v17i1.9015

Muhammad Aurongzeb Jamil-ur-Rahman Center for Genome Research, Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, 75270, Pakistan
Yasmeen Rashid Department of Biochemistry, University of Karachi, Karachi, 75270, Pakistan
Syed Habib Ahmed Naqvi Institute of Biotechnology & Genetic Engineering, University of Sindh, Jamshoro, Pakistan
Ambrina Khatoon Department of Research, Ziauddin University, 4/B, Saharah-e-Ghalib, Block-6, Clifton, Karachi, 7500, Pakistan
Sadia Abdul Haq Department of Biochemistry, University of Karachi, Karachi, 75270, Pakistan
Muhammad Kamran Azim Department of Biosciences, Mohammad Ali Jinnah University, Karachi, Pakistan
Imdad Kaleem Department of Bioinformatics and Bioscience, COMSATS University (CUI), Islamabad, Pakistan
Shahid Bashir Neuroscience Center, King Fahad Specialist Hospital, Dammam, Saudi Arabia

DOI: https://doi.org/10.18502/ijpa.v17i1.9015

Keywords: Naegleria fowleri; Genotyping; Primary amoebic menin-goencephalitis

Abstract

Background: Primary amoebic meningoencephalitis (PAM) is an acute and fulminant CNS infection caused by Naegleria fowleri. Recreational activities and ritual ablution with contaminated warm fresh water are the main reason of PAM. Pakistan ranked the second most affected country, where most of the PAM incidences were reported from Karachi, Pakistan.

Methods: In May, 2019, a 28-yr-old suspected PAM patient came to the Imam Zain-Ul-Abdin Hospital, Karachi. Biochemical and cytological investigations of patient`s CSF were carried out at Karachi Diagnostic Center and Molecular Biology Lab. Sequencing of Naegleria sp. specific (ITS) primer-based amplicons was performed from both patient`s CSF and water samples followed by multiple sequence alignment and phylogenetic studies.

Results: Biochemical and cytological investigations of patient`s CSF showed 5 mg/dl glucose, 240 mg/dl total protein and 2260/mm³TLC suggesting acute meningoencephalitis. PCR-based analyses of patient`s CSF and his residential tap water samples using Naegleria sp. specific (ITS) and N. fowleri specific primers revealed the presence of N. fowleri DNA. Nucleotide sequences of ITS primer-based amplicons from both patient`s CSF and water samples were submitted in GenBank under the accession numbers MT726981.1 and MT726226.1, respectively. According to phylogenetic analysis, N. fowleri isolate from Pakistan has shown the least node age of seven.

Conclusion: Here, for the very first time in Pakistan, N. fowleri genotype has been identified as type-2. Phylogenetic analysis showed that N. fowleri isolate from Pakistan is among the latest descendants, i.e., evolved later in life.